Transcriptome Sequencing of Codonopsis pilosula and Identification of Candidate Genes Involved in Polysaccharide Biosynthesis

被引:36
作者
Gao, Jian Ping [1 ]
Wang, Dong [1 ]
Cao, Ling Ya [1 ]
Sun, Hai Feng [2 ]
机构
[1] Shanxi Med Univ, Coll Pharm, Taiyuan, Shanxi, Peoples R China
[2] Shanxi Univ, Coll Chem & Chem Engn, Taiyuan, Shanxi, Peoples R China
基金
中国国家自然科学基金;
关键词
EXPRESSION; TOOL; GLYCOSYLTRANSFERASES; CLONING; ROOTS; PCR;
D O I
10.1371/journal.pone.0117342
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Background Codonopsis pilosula (Franch.) Nannf. is one of the most widely used medicinal plants. Although chemical and pharmacological studies have shown that codonopsis polysaccharides (CPPs) are bioactive compounds and that their composition is variable, their biosynthetic pathways remain largely unknown. Next-generation sequencing is an efficient and high-throughput technique that allows the identification of candidate genes involved in secondary metabolism. Principal Findings To identify the components involved in CPP biosynthesis, a transcriptome library, prepared using root and other tissues, was assembled with the help of Illumina sequencing. A total of 9.2 Gb of clean nucleotides was obtained comprising 91,175,044 clean reads, 102,125 contigs, and 45,511 unigenes. After aligning the sequences to the public protein databases, 76.1% of the unigenes were annotated. Among these annotated unigenes, 26,189 were assigned to Gene Ontology categories, 11,415 to Clusters of Orthologous Groups, and 18,848 to Kyoto Encyclopedia of Genes and Genomes pathways. Analysis of abundance of transcripts in the library showed that genes, including those encoding metallothionein, aquaporin, and cysteine protease that are related to stress responses, were in the top list. Among genes involved in the biosynthesis of CPP, those responsible for the synthesis of UDP-L-arabinose and UDP-xylose were highly expressed. Significance To our knowledge, this is the first study to provide a public transcriptome dataset prepared from C. pilosula and an outline of the biosynthetic pathway of polysaccharides in a medicinal plant. Identified candidate genes involved in CPP biosynthesis provide understanding of the biosynthesis and regulation of CPP at the molecular level.
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页数:25
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共 56 条
[31]  
Rozen S, 2000, Methods Mol Biol, V132, P365
[32]   Analyzing real-time PCR data by the comparative CT method [J].
Schmittgen, Thomas D. ;
Livak, Kenneth J. .
NATURE PROTOCOLS, 2008, 3 (06) :1101-1108
[33]   Immunological Adjuvant Effect of a Water-Soluble Polysaccharide, CPP, from the Roots of Codonopsis pilosula on the Immune Responses to Ovalbumin in Mice [J].
Sun, Yong-Xu .
CHEMISTRY & BIODIVERSITY, 2009, 6 (06) :890-896
[34]   MEGA5: Molecular Evolutionary Genetics Analysis Using Maximum Likelihood, Evolutionary Distance, and Maximum Parsimony Methods [J].
Tamura, Koichiro ;
Peterson, Daniel ;
Peterson, Nicholas ;
Stecher, Glen ;
Nei, Masatoshi ;
Kumar, Sudhir .
MOLECULAR BIOLOGY AND EVOLUTION, 2011, 28 (10) :2731-2739
[35]   An efficient approach to finding Siraitia grosvenorii triterpene biosynthetic genes by RNA-seq and digital gene expression analysis [J].
Tang, Qi ;
Ma, Xiaojun ;
Mo, Changming ;
Wilson, Iain W. ;
Song, Cai ;
Zhao, Huan ;
Yang, Yanfang ;
Fu, Wei ;
Qiu, Deyou .
BMC GENOMICS, 2011, 12
[36]   The COG database: an updated version includes eukaryotes [J].
Tatusov, RL ;
Fedorova, ND ;
Jackson, JD ;
Jacobs, AR ;
Kiryutin, B ;
Koonin, EV ;
Krylov, DM ;
Mazumder, R ;
Mekhedov, SL ;
Nikolskaya, AN ;
Rao, BS ;
Smirnov, S ;
Sverdlov, AV ;
Vasudevan, S ;
Wolf, YI ;
Yin, JJ ;
Natale, DA .
BMC BIOINFORMATICS, 2003, 4 (1)
[37]   Dung-shen (Codonopsis pilosula) attenuated the cardiac-impaired insulin-like growth factor II receptor pathway on myocardial cells [J].
Tsai, Kun-Hsi ;
Lee, Nien-Hung ;
Chen, Guei-Ying ;
Hu, Wei-Syun ;
Tsai, Chen-Yen ;
Chang, Mu-Hsin ;
Jong, Gwo-Ping ;
Kuo, Chia-Hua ;
Tzang, Bor-Show ;
Tsai, Fuu-Jen ;
Tsai, Chang-Hai ;
Huang, Chih-Yang .
FOOD CHEMISTRY, 2013, 138 (2-3) :1856-1867
[38]   Cloning and endogenous expression of a Eucalyptus grandis UDP-glucose dehydrogenase cDNA [J].
Veneziano Labate, Monica T. ;
Ferreira Bertolo, Ana L. ;
do Nascimento, Daniela Defavari ;
Gutmanis, Gunta ;
de Andrade, Alexander ;
Calderan Rodrigues, Maria J. ;
Camargo, Eduardo L. O. ;
Boaretto, Luis Felipe ;
Moon, David H. ;
Bragatto, Juliano ;
Labate, Carlos A. .
GENETICS AND MOLECULAR BIOLOGY, 2010, 33 (04) :686-U131
[39]   Overexpression of a cytosol-localized rhamnose biosynthesis protein encoded by Arabidopsis RHM1 gene increases rhamnose content in cell wall [J].
Wang, Jinfeng ;
Ji, Qinmei ;
Jiang, Ling ;
Shen, Songdong ;
Fan, Yunliu ;
Zhang, Chunyi .
PLANT PHYSIOLOGY AND BIOCHEMISTRY, 2009, 47 (02) :86-93
[40]   Structure, mechanism and engineering of plant natural product glycosyltransferases [J].
Wang, Xiaoqiang .
FEBS LETTERS, 2009, 583 (20) :3303-3309